Information on EC 3.4.22.32 - Stem bromelain

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The expected taxonomic range for this enzyme is: Ananas comosus

EC NUMBER
COMMENTARY
3.4.22.32
-
RECOMMENDED NAME
GeneOntology No.
Stem bromelain
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
broad specificity for cleavage of proteins, but strong preference for Z-Arg-Arg-/-NHMec amongst small molecule substrates
show the reaction diagram
; endopeptidase; The most abundant of the cysteine endopeptidases of the stem of the pineapple plant, Ananas comosus. Distinct from the bromelain found in the pineapple fruit (3.4.22.33). Scarcely inhibited by chicken cystatin and also very slowly inactivated by 3.4.22.4.
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acidic bromelain stem proteinase
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Bromelain
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-
-
-
Bromelain, stem
-
-
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EC 3.4.22.4
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formerly
-
EC 3.4.4.24
-
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formerly
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Pineapple stem bromelain
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-
-
-
Pineapple stem bromelain
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-
CAS REGISTRY NUMBER
COMMENTARY
37189-34-7
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
2 enzyme forms: pI 9.45 (basic form), pI 4.7 (acidic form)
-
-
Manually annotated by BRENDA team
2 forms: SB1 and SB2
-
-
Manually annotated by BRENDA team
2 isozymes SBA/a and SBA/b
-
-
Manually annotated by BRENDA team
6 enzyme forms: SBB 1-5 and SBA
-
-
Manually annotated by BRENDA team
at least 8 basic proteolytically active components
-
-
Manually annotated by BRENDA team
bromelain A and B
-
-
Manually annotated by BRENDA team
enzyme forms: I-1, I-2, II-1, II-2
-
-
Manually annotated by BRENDA team
purified enzyme
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
bromelain increases expression of p53 as well as Bax in mouse skin papillomas, at the same time, bromelain decreases the activity of cell survival regulators such as Akt and Erk thus promoting apoptotic cell death in tumors. Bromelain reduces CD44 on the surface of mouse and human tumor cells accompanied by diminished cancer cell invasion and substrate attachment as well as by attenuation of de novo protein synthesis. Bromelain can stimulate the innate immune system by activating neutrophils to produce reactive oxygen species
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
azocasein + H2O
fragments of azocasein
show the reaction diagram
-
-
-
-
Azocoll + H2O
Hydrolyzed azocoll
show the reaction diagram
-
-
-
-
Benzoyl-Arg-Arg 4-nitrophenyl ester + H2O
Benzoyl-Arg-Arg + 4-nitrophenol
show the reaction diagram
-
-
-
-
-
Benzoyl-citrulline 4-nitroanilide + H2O
Benzoyl-citrulline + 4-nitroaniline
show the reaction diagram
-
-
-
-
-
Benzoyl-DL-Ala ethyl ester + H2O
Benzoyl-DL-Ala + ethanol
show the reaction diagram
-
-
-
-
-
Benzoyl-DL-Arg 4-nitroanilide + H2O
Benzoyl-DL-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
-
Benzoyl-Gly ethyl ester + H2O
Benzoyl-Gly + ethanol
show the reaction diagram
-
-
-
-
-
Benzoyl-L-Arg amide + H2O
Benzoyl-L-Arg + NH3
show the reaction diagram
-
-
-
-
-
Benzoyl-L-Arg ethyl ester + H2O
Benzoyl-L-Arg + ethanol
show the reaction diagram
-
-
-
-
-
Benzoyl-L-Arg methyl ester + H2O
Benzoyl-L-Arg + methanol
show the reaction diagram
-
-
-
-
-
Benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
convenient substrate for stem bromelain, scarcely affected by fruit bromelain
-
-
?
Benzyloxycarbonyl-Arg-Arg 4-nitrophenyl ester + H2O
Benzyloxycarbonyl-Arg-Arg + 4-nitrophenol
show the reaction diagram
-
-
-
-
-
benzyloxycarbonyl-Arg-Arg-NH-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
synthetic substrate
-
?
benzyloxycarbonyl-Arg-Arg-p-nitroanilide + H2O
benzyloxycarbonyl-Arg-Arg + p-nitroanilide
show the reaction diagram
-
-
-
-
?
Benzyloxycarbonyl-citrulline 4-nitroanilide + H2O
Benzyloxycarbonyl-citrulline + 4-nitroaniline
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly 4-nitrophenyl ester + H2O
Benzyloxycarbonyl-Gly + 4-nitrophenol
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-citrulline 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-Phe-citrulline 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-Phe-citrulline 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Gly-Phe-Phe-citrulline 4-nitroanilide + H2O
?
show the reaction diagram
-
best substrate
-
-
-
Benzyloxycarbonyl-Gly-Pro-citrulline 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
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Benzyloxycarbonyl-L-Ala 4-nitrophenyl ester + H2O
Benzyloxycarbonyl-L-Ala + 4-nitrophenol
show the reaction diagram
-
-
-
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Benzyloxycarbonyl-L-Asn 4-nitrophenyl ester + H2O
Benzyloxycarbonyl-L-Asn + 4-nitrophenol
show the reaction diagram
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-
-
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Benzyloxycarbonyl-L-Lys 4-nitrophenyl ester + H2O
Benzyloxycarbonyl-L-Lys + 4-nitrophenol
show the reaction diagram
-
-
-
-
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benzyloxycarbonyl-L-Tyr 4-nitrophenyl ester + H2O
benzyloxycarbonyl-L-Tyr + 4-nitrophenol
show the reaction diagram
-
-
-
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-
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide + H2O
Benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
-
benzyloxycarbonyl-Phe-Arg-NH-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
synthetic substrate
-
?
Benzyloxycarbonyl-Phe-citrulline 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
benzyloxycarbonyl-Phe-Val-Arg-NH-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
synthetic substrate
-
?
Benzyloxycarbonyl-Pro-citrulline 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
Bz-Phe-Val-Arg-4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
hydrolyzed casein
show the reaction diagram
-
-
-
-
-
casein + H2O
hydrolyzed casein
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
P14518
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-
-
?
casein + H2O
L-Tyr + ?
show the reaction diagram
-
-
-
-
?
CD25 + H2O
?
show the reaction diagram
-
bromelain proteolytically cleaved cell-surface CD25 from activated CD4+ T cells, a mechanism of action to exert therapeutic benefits in inflammatory conditions, overview, soluble CD25is a therapeutic target in inflammation, autoimmunity and allergy
-
-
?
Fibrin + H2O
Hydrolyzed fibrin
show the reaction diagram
-
-
-
-
Gelatin + H2O
?
show the reaction diagram
-
-
-
-
?
Gly ethyl ester + H2O
Gly + ethanol
show the reaction diagram
-
-
-
-
-
Hemoglobin + H2O
Hydrolyzed hemoglobin
show the reaction diagram
-
-
-
-
Hemoglobin + H2O
Hydrolyzed hemoglobin
show the reaction diagram
-
denatured
-
-
-
hemoglobin + H2O
L-Tyr + ?
show the reaction diagram
-
-
-
-
?
Hide powder + H2O
Hydrolyzed hide powder
show the reaction diagram
-
-
-
-
Isolated soybean protein + H2O
?
show the reaction diagram
-
-
-
-
-
L-Phe ethyl ester + H2O
L-Phe + ethanol
show the reaction diagram
-
-
-
-
-
L-Pyroglutamyl-L-Phe-L-Leu 4-nitroanilide + H2O
?
show the reaction diagram
-
-
-
-
-
L-Tyr ethyl ester + H2O
L-Tyr + ethanol
show the reaction diagram
-
-
-
-
-
N-alpha-benzyloxycarbonyl-Lys-p-nitrophenyl ester + H2O
N-alpha-benzyloxycarbonyl-Lys + p-nitrophenol
show the reaction diagram
-
-
-
-
?
N-Benzyloxycarbonyl-L-Phe-L-Ser methyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
Nalpha-CBZ-L-lysine 4-nitrophenyl ester + H2O
Nalpha-CBZ-L-lysine + 4-nitrophenol
show the reaction diagram
-
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
Pyr-Phe-Lys-4-nitroanilide + H2O
Pyr-Phe-Lys + 4-nitroaniline
show the reaction diagram
-
isozyme SBA/b is about 3fold less active than isozyme SBA/a
-
?
tert-butyloxycarbonyl-Leu-Arg-Arg-4-methylcoumaryl-7-amide + H2O
tert-butyloxycarbonyl-Leu-Arg-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
Tosyl-citrulline 4-nitroanilide + H2O
Tosyl-citrulline + 4-nitroaniline
show the reaction diagram
-
-
-
-
-
Tosyl-L-Arg methyl ester + H2O
Tosyl-L-Arg + methanol
show the reaction diagram
-
-
-
-
-
Tosyl-L-Lys methyl ester + H2O
Tosyl-L-Lys + methanol
show the reaction diagram
-
-
-
-
-
Wheat gluten + H2O
Hydrolyzed wheat gluten
show the reaction diagram
-
-
-
-
-
Z-Arg-Arg-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
-
-
-
-
?
Meuchenia sp. insoluble muscle protein + H2O
hydrolyzed Meuchenia sp. insoluble muscle protein
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
-
-
-
-
additional information
?
-
-
overview: synthetic substrates
-
-
-
additional information
?
-
-
substrate conformation of N-acylglycine thioester substrates in the active site by resonance spectroscopy
-
-
-
additional information
?
-
-
enzyme reduces the amount of prostaglandin-E2 and substance P in vivo in inflammatory process in rats, while in vitro it induces an increase in the concentration of these inflammation mediators, thus the enzyme does not directly interact with the compounds, but has an antiinflammatory effect in vivo
-
?
additional information
?
-
-
bromelain from pineapple stem shows therapeutic benefits in a variety of inflammatory diseases, including murine inflammatory bowel disease, mechanism, overview. Bromelain primary long-term effect is abrogation of firm adhesion of leukocytes to blood vessels at the site of inflammation. These changes in adhesion are correlated with rapid re-expression of the bromelain-sensitive CD62L/L-selectin molecules that mediate rolling following in vivo bromelain treatment and minimal re-expression of CD128
-
-
-
additional information
?
-
-
bromelain possesses anti-inflammatory activity and reduces blood viscosity, prevents the aggregation of blood platelets, and improves ischemia-reperfusion injury, I/R, in a skeletal muscle model in adult Sprague-Dawley rats. The enzyme increases phosphorylation of Akt in rat heart both in the cytosolic and the nuclear fraction following I/R
-
-
-
additional information
?
-
-
CD4+ T cells retain the ability to divide after bromelain treatment
-
-
-
additional information
?
-
-
effects of orally administered bromelain in an ovalbumin-induced murine model of acute allergic airway disease in female C57BL/6J mice, bromelain causes decreased methacholine sensitivity, reduction in bronchoalveolar lavage eosinophils and interleukin-13 concentrations, as well as of CD19+ B cells and CD8+ T cells, as compared with controls, overview. Bromelain modulates lung lymphocytes during acute asthma
-
-
-
additional information
?
-
-
phosphorylation and consequent degradation of IkappaBalpha, the inhibitor of the nuclear importing sequences of NF-kappaB, is blocked by bromelain, which also shows anti-inflammatory, anti-invasive and anti-metastatic properties, anti-tumor-initiating effects of bromelain in 2-stage mouse skin tumorigenesis model, overview. Bromelain treatment resulted in upregulation of p53 and Bax and subsequent activation of caspase 3 and caspase 9 with concomitant decrease in Bcl-2, and bromelain inhibits DMBA-induced DNA alkylation damage, overview
-
-
-
additional information
?
-
-
bromelain proteolytically removes in vitro a number of cell surface molecules that are vital to leukocyte trafficking, including CD128a/CXCR1 and CD128b/CXCR2 that serve as receptors for the human neutrophil chemoattractant IL-8 and its murine homologues. In vivo bromelain treatment generates a 50-85% reduction in human neutrophil migration in 3 different murine models of leukocyte migration into the inflamed peritoneal cavity, and bromelain treatment inhibits IL-8-stimulated migration of human neutrophils in vitro
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
protein + H2O
peptides
show the reaction diagram
-
-
-
?
CD25 + H2O
?
show the reaction diagram
-
bromelain proteolytically cleaved cell-surface CD25 from activated CD4+ T cells, a mechanism of action to exert therapeutic benefits in inflammatory conditions, overview
-
-
?
additional information
?
-
-
enzyme reduces the amount of prostaglandin-E2 and substance P in vivo in inflammatory process in rats, while in vitro it induces an increase in the concentration of these inflammation mediators, thus the enzyme does not directly interact with the compounds, but has an antiinflammatory effect in vivo
-
?
additional information
?
-
-
bromelain from pineapple stem shows therapeutic benefits in a variety of inflammatory diseases, including murine inflammatory bowel disease, mechanism, overview. Bromelain primary long-term effect is abrogation of firm adhesion of leukocytes to blood vessels at the site of inflammation. These changes in adhesion are correlated with rapid re-expression of the bromelain-sensitive CD62L/L-selectin molecules that mediate rolling following in vivo bromelain treatment and minimal re-expression of CD128
-
-
-
additional information
?
-
-
bromelain possesses anti-inflammatory activity and reduces blood viscosity, prevents the aggregation of blood platelets, and improves ischemia-reperfusion injury, I/R, in a skeletal muscle model in adult Sprague-Dawley rats. The enzyme increases phosphorylation of Akt in rat heart both in the cytosolic and the nuclear fraction following I/R
-
-
-
additional information
?
-
-
CD4+ T cells retain the ability to divide after bromelain treatment
-
-
-
additional information
?
-
-
effects of orally administered bromelain in an ovalbumin-induced murine model of acute allergic airway disease in female C57BL/6J mice, bromelain causes decreased methacholine sensitivity, reduction in bronchoalveolar lavage eosinophils and interleukin-13 concentrations, as well as of CD19+ B cells and CD8+ T cells, as compared with controls, overview. Bromelain modulates lung lymphocytes during acute asthma
-
-
-
additional information
?
-
-
phosphorylation and consequent degradation of IkappaBalpha, the inhibitor of the nuclear importing sequences of NF-kappaB, is blocked by bromelain, which also shows anti-inflammatory, anti-invasive and anti-metastatic properties, anti-tumor-initiating effects of bromelain in 2-stage mouse skin tumorigenesis model, overview. Bromelain treatment resulted in upregulation of p53 and Bax and subsequent activation of caspase 3 and caspase 9 with concomitant decrease in Bcl-2, and bromelain inhibits DMBA-induced DNA alkylation damage, overview
-
-
-
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-hydroxyethyl disulfide
-
-
alpha2-Macroglobulin
-
-
-
Ananas comosus bromelain inhibitor
-
competitive inhibition, several isoforms of the native double-chain protein, inhibition mechanism, structure-function relationship, structure modeling of inhibitor protein, bromelain inhibitor VI: PDB 1Bl6
-
Ananas comosus bromelain inhibitor
-
purified from stem, several isoforms of the native inhibitor protein consisting of multiple light and heavy chains, overview, 3D-structure model
-
Cystatin
-
chicken, weak inhibition
-
E-64
-
weak inhibition
ethanol
-
competitive inhibitor
Guanidine-HCl
-
complete loss of activity at 6 M guanidine-HCl
Human plasma
-
-
-
Inhibitor from pineapple stem
-
properties of the inhibitor
-
Inhibitor from pineapple stem
-
-
-
iodoacetic acid
-
-
PCMB
-
cysteine restores activity
phenylmercuric acetate
-
-
Potato cysteine proteinase inhibitor
-
-
-
Rabbit anti-stem bromelain antibodies
-
-
-
SDS
-
SDS acts as a partial non-competitive inhibitor, 97% inhibition at 100 mM, detailed interaction study and inhibition mechanism, bromelain is partly resistant to SDS binding and denaturation, overview. SDS at submicellar level, below 5 mM, causes conformation change of bromelain leading to a stable entity
SDS
-
for native bromelain, SDS at 4 and 5 mg/ml causes a significant inhibition of 57% and 61%, respectively
Sucrose
-
bromelain in the presence of 1 M sucrose is destabilized under thermal stress, the average melting temperature decrease by 5C, additionally the enzyme is inactivated faster at 60C
Sulfhydryl reagent
-
-
-
sulfur dioxide
-
strong inhibitor
-
trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane
-
i.e. E-64, slow inactivation
trehalose
-
bromelain in the presence of 1 M trehalose is destabilized under thermal stress, the average melting temperature decrease by 7C, additionally the enzyme is inactivated faster at 60C
L-3-carboxy-2,3-trans-epoxypropionylleucylamido(4-guanidino)butane
-
irreversible
additional information
-
not: DFP
-
additional information
-
alpha1-antitrypsin
-
additional information
-
scarcely inhibited by chicken cystatin (amino acid sequence and implications for weak binding of cystatin)
-
additional information
-
no inhibition by cystatin
-
additional information
-
reversible inactivation of the protease in 10 mM Na-phosphate, pH 7.5 containing 5 mM of Na-tetrathionate, during purification, reactivation by 8 mM of DTT and 4 mM of EDTA in the same buffer at 25C
-
additional information
-
gallic tannins or ellagic tannins show no inhibitory activity
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2,3-Dimercapto-1-propanol
-
not as effective in activation as cysteine
cysteine
-
inactive unless treated with
DTT
-
not as effective in activation as cysteine
Thioglycol
-
not as effective in activation as cysteine
Urea
-
at 5 M and 8-9 M
Guanidine-HCl
-
at 2 mM
additional information
-
like ficin and unlike papain, bromelain does not possess a carboxyl group capable of enhancing the rate of reaction of the active-centre thiol group with 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole
-
additional information
-
bromelain activator solution consists of 0.02 M EDTA and 0.05 M L-cysteine, pH 8.0
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
210
-
Benzoyl-Gly ethyl ester
-
-
1.2
-
Benzoyl-L-Arg amide
-
-
3.1
-
Benzoyl-L-Arg amide
-
enzyme form I-1
6
-
Benzoyl-L-Arg amide
-
enzyme form II-2
6.6
-
Benzoyl-L-Arg amide
-
enzyme form II-1
9.5
-
Benzoyl-L-Arg amide
-
enzyme form I-2
67
-
Benzoyl-L-Arg amide
-
acidic enzyme form
57
-
benzoyl-L-Arg ethyl ester
-
acidic enzyme form
170
-
benzoyl-L-Arg ethyl ester
-
-
190
-
benzoyl-L-Arg ethyl ester
-
enzyme form SB1
200
-
benzoyl-L-Arg ethyl ester
-
enzyme form SB2, benzoyl-Gly ethyl ester, enzyme form SB1 and SB2
32
-
benzoyl-L-Arg methyl ester
-
-
0.0097
-
benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide
-
-
0.0154
-
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
-
-
0.0074
-
benzyloxycarbonyl-Arg-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/b, pH 6.0, 25C
0.0081
-
benzyloxycarbonyl-Arg-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/a, pH 6.0, 25C
0.0153
-
Benzyloxycarbonyl-Gly-Phe-citrulline 4-methylcoumarin 7-amide
-
-
0.0831
-
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
-
-
0.0554
-
benzyloxycarbonyl-Phe-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/a, pH 6.0, 25C
0.0636
-
benzyloxycarbonyl-Phe-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/b, pH 6.0, 25C
0.003
-
benzyloxycarbonyl-Phe-Val-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/b, pH 6.0, 25C
0.0039
-
benzyloxycarbonyl-Phe-Val-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/a, pH 6.0, 25C
0.04
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, free SO2 25 mg/l
-
0.052
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, seed 1.2 g/l
-
0.056
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, free SO2 10 mg/l
-
0.079
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, skin 0.99 g/l
-
0.0952
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, seed 0.3 g/l
-
0.118
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ethanol 0%
-
0.152
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, skin 0.25 g/l
-
0.22
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, gallic 0.02 g/l
-
0.25
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ethanol 12%; pH 3.2, 25C, without free SO2; pH 3.2, 25C, without gallic; pH 3.2, 25C, without skin
-
0.2558
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ellagic 0.02 g/l
-
0.27
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ellagic 0.04 g/l; pH 3.2, 25C, gallic 0.03 g/l
-
0.305
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ethanol 18%
-
0.129
-
casein
-
native enzyme, at pH 7.0
0.217
-
casein
-
native enzyme, in the presence of 5 M urea at pH 7.0
0.221
-
casein
-
native enzyme, in the presence of 2 M guanidine-HCl at pH 7.0
2.83
-
casein
-
poly(maleic anhydride)-modified enzyme, in 50 mM Tris-HCl buffer, pH 8.0, at 40C
4.09
-
casein
-
pyromellitic anhydride-modified enzyme, in 50 mM Tris-HCl buffer, pH 8.0, at 40C
5.26
-
casein
-
native enzyme, in 50 mM Tris-HCl buffer, pH 8.0, at 40C
0.4
-
L-Pyroglutamyl-L-Phe-L-Leu 4-nitroanilide
-
enzyme form F9
2.3
-
L-Pyroglutamyl-L-Phe-L-Leu 4-nitroanilide
-
enzyme form F4
0.53
-
N-Benzyloxycarbonyl-L-Phe-L-Ser methyl ester
-
-
84
-
tosyl-L-Lys ethyl ester
-
-
2.42
-
L-Pyroglutamyl-L-Phe-L-Leu 4-nitroanilide
-
enzyme form F5
additional information
-
additional information
-
pH-dependence of Km
-
additional information
-
additional information
-
kinetics of hydrolysis and influence of modifiers of hydrolysis of N-benzoyl-L-Ser methyl ester
-
additional information
-
additional information
-
kinetics of reaction of benzofuroxan, a thiol-specific reactivity probe with the enzyme
-
additional information
-
additional information
-
casein: Km 0.1%
-
additional information
-
additional information
-
Km-for casein is 1.1 mg/ml for the native enzyme, 2 mg/ml for the enzyme covalently coupled to the CNBr-activated Sepharose and 0.54 mg/ml for enzyme affinity-bound to a Sepharose matrix precoupled with the lactin concanavalin A
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.55
-
acetyl-Gly ethyl ester
-
-
0.21
-
Benzoyl-Gly ethyl ester
-
enzyme form SB1
0.24
-
Benzoyl-Gly ethyl ester
-
enzyme form SB2
0.36
-
Benzoyl-Gly ethyl ester
-
-
0.0035
-
Benzoyl-L-Arg amide
-
-
0.0039
-
Benzoyl-L-Arg amide
-
enzyme form I-2
0.0046
-
Benzoyl-L-Arg amide
-
enzyme form I-1
0.018
-
Benzoyl-L-Arg amide
-
enzyme form II-2
0.027
-
Benzoyl-L-Arg amide
-
enzyme form II-1
0.39
-
benzoyl-L-Arg ethyl ester
-
enzyme form SB2
0.42
-
benzoyl-L-Arg ethyl ester
-
enzyme form SB1
0.5
-
benzoyl-L-Arg ethyl ester
-
-
0.11
-
benzoyl-L-Arg methyl ester
-
-
0.893
-
benzoyl-Phe-Val-Arg 4-methylcoumarin 7-amide
-
-
27
-
benzyloxycarbonyl-Arg-Arg 4-methylcoumarin 7-amide
-
-
0.024
-
benzyloxycarbonyl-Arg-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/a, pH 6.0, 25C
0.04
-
benzyloxycarbonyl-Arg-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/b, pH 6.0, 25C
1.75
-
Benzyloxycarbonyl-Gly 4-nitrophenyl ester
-
bromelain A
3.35
-
Benzyloxycarbonyl-Gly 4-nitrophenyl ester
-
bromelain A
0.12
-
Benzyloxycarbonyl-Gly-Phe-citrulline 4-methylcoumarin 7-amide
-
-
2.5
-
benzyloxycarbonyl-L-Ala 4-nitrophenyl ester
-
bromelain A
8.3
-
benzyloxycarbonyl-L-Ala 4-nitrophenyl ester
-
bromelain B
1.4
1.5
Benzyloxycarbonyl-L-Asn 4-nitrophenyl ester
-
-
7.4
-
benzyloxycarbonyl-L-Lys 4-nitrophenyl ester
-
bromelain A
9.8
-
benzyloxycarbonyl-L-Lys 4-nitrophenyl ester
-
bromelain B
0.4
-
benzyloxycarbonyl-L-Tyr 4-nitrophenyl ester
-
bromelain A
0.44
-
benzyloxycarbonyl-L-Tyr 4-nitrophenyl ester
-
bromelain B
0.13
-
Benzyloxycarbonyl-Phe-Arg 4-methylcoumarin 7-amide
-
-
0.49
-
benzyloxycarbonyl-Phe-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/a, pH 6.0, 25C
0.56
-
benzyloxycarbonyl-Phe-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/b, pH 6.0, 25C
6.08
-
benzyloxycarbonyl-Phe-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/b, pH 6.0, 25C
5.1
-
benzyloxycarbonyl-Phe-Val-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/a, pH 6.0, 25C
6.4
-
benzyloxycarbonyl-Phe-Val-Arg-NH-4-methylcoumarin 7-amide
-
isozyme SBA/b, pH 6.0, 25C
0.39
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, free SO2 25 mg/l
-
0.96
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, free SO2 10 mg/l
-
4.32
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, seed 1.2 g/l
-
6.31
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, skin 0.99 g/l
-
6.55
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, seed 0.3 g/l
-
8.44
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, skin 0.25 g/l
-
16.94
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, gallic 0.03 g/l
-
18.33
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ellagic 0.04 g/l
-
18.35
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, gallic 0.02 g/l
-
19.23
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ellagic 0.02 g/l
-
19.43
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ethanol 0%
-
20.03
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ethanol 12%; pH 3.2, 25C, without free SO2; pH 3.2, 25C, without gallic; pH 3.2, 25C, without skin
-
21.45
-
Bz-Phe-Val-Arg-4-nitroanilide
-
pH 3.2, 25C, ethanol 18%
-
0.41
-
casein
-
native enzyme, at pH 7.0
0.97
-
casein
-
native enzyme, in the presence of 5 M urea at pH 7.0
0.99
-
casein
-
native enzyme, in the presence of 2 M guanidine-HCl at pH 7.0
11.72
-
casein
-
native enzyme, at pH 7.0
17.33
-
casein
-
native enzyme, in the presence of 5 M urea at pH 7.0
18.08
-
casein
-
native enzyme, in the presence of 2 M guanidine-HCl at pH 7.0
0.68
-
L-Pyroglutamyl-L-Phe-L-Leu 4-nitroanilide
-
enzyme form F5
0.87
-
L-Pyroglutamyl-L-Phe-L-Leu 4-nitroanilide
-
enzyme form F4
3.94
-
L-Pyroglutamyl-L-Phe-L-Leu 4-nitroanilide
-
enzyme form F9
3.4
-
N-Benzyloxycarbonyl-L-Phe-L-Ser methyl ester
-
-
0.035
-
Tosyl-L-Lys methyl ester
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.145
-
SDS
-
25C
0.1
-
Chicken cystatin
-
both isozymes, pH 6.0, 25C
-
additional information
-
additional information
-
k2/Ki for E-64
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.55
-
SDS
-
25C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.17
-
-
partially purified enzyme, 37C, pH 6.5
0.62
-
-
purified enzyme, substrate casein
additional information
-
-
assay methods
additional information
-
-
-
additional information
-
-
activity of glycosylated and deglycosylated enzyme at different temperatures and pH 7.0, and at different pH-values and 37C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3.2
-
-
assay at
4
4.5
-
L-pyroglutamyl-L-Phe-L-Leu 4 nitroanilide, enzyme form F4 and F5
4.6
-
-
assay at
5
8
-
wide plateau, Nalpha-benzoyl-L-Arg amide
5
-
-
denatured hemoglobin, enzyme form F4
6
7
-
substrate-dependent
6
-
-
immobilized enzyme
7
8.5
-
casein
7
-
-
about, L-pyroglutamyl-L-Phe-L-Leu 4-nitroanilide, enzyme form F9
7
-
-
assay at
7
-
-
native enzyme
7.5
-
-
-
7.5
-
-
native enzyme, enzyme covalently coupled to the CNBr-activated Sepharose and enzyme affinity-bound to a Sepharose matrix precoupled with the lactin concanavalin A
7.5
-
-
native enzyme
8
-
-
native enzyme, at 35C
10
-
-
randomly polymer-coupled enzyme preparations, at 35C
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
10
-
5: about 40% of activity maximum, 10.0: about 70% of activity maximum
5
11
-
pH 5.0: native enzyme shows about 25% of maximal activity, enzyme covalently coupled to the CNBr-activated Sepharose shows about 30% of maximal activity, enzyme affinity-bound to a Sepharose matrix precoupled with the lactin concanavalin A shows about 35% of maximal activity, pH 11: native enzyme shows about 25% of maximal activity, enzyme covalently coupled to the CNBr-activated Sepharose shows about 30% of maximal activity, enzyme affinity-bound to a Sepharose matrix precoupled with the lactin concanavalin A shows about 60% of maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at
60
70
-
randomly polymer-coupled enzyme preparations
60
-
-
native enzyme
60
-
P14518
-
62.5
-
-
-
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.8
-
-
isoelectric focusing, isozyme SBA/b, and isozyme SBA/a variant 2
4.9
-
-
isoelectric focusing, isozyme SBA/a variant 1
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
18000
-
-
Ananas comosus, enzyme form I-2, II-1 and II-2, gel filtration
18800
-
-
Ananas comosus, bromelain A, gel filtration
19700
-
-
Ananas comosus, bromelain B, gel filtration
22500
-
-
Ananas comosus, gel filtration
23430
-
-
Ananas comosus, enzyme form F9, mass spectroscopy
23550
-
-
isozyme SBA7A, mass spectroscopy
23560
-
-
isozyme SBA/b, mass spectroscopy
23800
-
-
SDS-PAGE
24400
-
-
Ananas comosus, enzyme form F4, mass spectroscopy
24470
-
-
Ananas comosus, enzyme form F5, mass spectroscopy
26000
-
-
SDS-PAGE
28000
-
-
Ananas comosus, enzyme form I-1, gel filtration
28000
-
-
Ananas comosus, enzyme form SB1 and SB2, sedimentation equilibrium method
28400
-
-
Ananas comosus, sedimentation velocity and equilibrium ultracentrifugation experiments
33000
-
-
Ananas comosus, determination from sedimentation and diffusion coefficient
35300
-
-
Ananas comosus, sedimentation velocity measurements
37000
-
-
Ananas comosus, gel filtration
43000
-
-
SDS-PAGE, recombinant protein
additional information
-
-
amino acid sequence
additional information
-
-
isolation and alignment of 3 peptides obtained by means of cyanogen bromide cleavage
additional information
-
-
amino acid sequence of the carboxyterminal-half
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 26000, Ananas comosus, SDS-PAGE
?
-
x * 23000, Ananas comosus, enzyme forms SBB 4 and 5, SBA; x * 27000, Ananas comosus, enzyme forms SBB 1-3, SDS-PAGE
?
-
x * 35000, Ananas comosus, bromelain A, SDS-PAGE
?
-
x * 25600, Ananas comosus, enzyme forms SB1 and SB2, SDS-PAGE
?
-
x * 23000, Ananas comosus, SDS-PAGE, 2 chains of MW 15000 and 8500 upon reduction with HS(CH2)2OH
monomer
-
1 * 28500, Ananas comosus, SDS-PAGE
monomer
-
1 * 37000, Ananas comosus
monomer
-
1 * 23550-27900, isozyme SBA/a, mass spectroscopy and SDS-PAGE, 1 * 23560-27800, isozyme SBA/b, mass spectroscopy and SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
-
glycoprotein
-
carbohydrate content: 4.3% (enzyme form SBB1), 3.6% (enzyme form SBB2), 3.5% (enzyme form SBB3), 8.2% (enzyme form SBA), enzyme form SBB4 and SBB5 contain no carbohydrate
glycoprotein
-
carbohydrate composition: molar ratio of mannose:xylose:fucose is 2.43:1.0:1.01 (enzyme form SB1), 2.55:1.17:0.92 (enzyme form SB2); neutral sugar content: 4.89% (enzyme form SB1), 5.24% (enzyme form SB2)
glycoprotein
-
enzyme form F4 and F5 contain fucose, N-acetylglucosamine, xylose and mannose in the ratio of 1.0:2.0:1.0:2.0, but only 50% of the proteins seem to be glycosylated, enzyme form F9 is unglycosylated
glycoprotein
-
N-glycosylated enzyme, glycosylation stabilizes the enzyme
glycoprotein
-
glycosylation stabilizes the enzyme, enzyme contains a single hetero-oligosaccharide unit per molecule, deglycosylated enzyme shows decreased activity
glycoprotein
-
both isozymes are highly glycosylated, isozyme SBA/a is about 2fold more glycosylated than isozyme SBA/b, glycoslyation composition
glycoprotein
-
contains a single oligosaccharide chain attached to the polypeptide
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.8
2
-
stem bromelain at pH 2.0 is maximally unfolded and characterized by significant loss of secondary structure (about 80%) and almost complete loss of tertiary contacts, at pH 0.8 a molten globule state is observed with secondary structure content similar to that of native protein but no tertiary structure
2
-
-
almost complete loss of native tertiary contacts, acid unfolded state
3
9
-
4C, 24 h, stable
4
10
-
5C, 24 h, stable
5
-
-
50C, 60 min, stable
5.5
10
-
stem bromelain is fully resistant against urea around neutral pH (5.5 to 10.0) and unfolds only below pH 5.0
5.6
9
-
stable
7
10
-
from pH 7.0 to 10.0, the protein's secondary structure remains the same, although a slight loss of tertiary structure is observed. Above pH 10.0, there is a significant and irreversible loss of secondary and tertiary structure. At pH 10.0, SBM shows a significant increase in 8-anilino-1-naphthalene-sulfonate binding relative to the native state. No significant loss of activity is observed up to pH 10.0, beyond which there is an irreversible loss of activity
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
recombinant protein stored at 4C shows a loss in enzymatic activity. The percentage loss in recombinant bromelain enzymatic activity is 27.5%
40
-
-
unstable above, crude enzyme extract
45
-
-
pH 7, 30 min, stable up to
45
-
-
stable up to in presence of casein
50
-
-
pH 5, 60 min, stable
55
-
-
pH 6.1, 20 min, 50% loss of activity
60
-
-
100 min, native enzyme loses 80% of initial activity, enzyme covalently coupled to the CNBr-activated Sepharose loses 70% of the initial activity, enzyme affinity-bound to a Sepharose matrix precoupled with the lactin concanavalin A loses 50% of initial activity
60
-
-
native bromelain rapidly loses activity at 60C and retains only about 10% of the initial activity after incubation for 180 min, about 70% and 58% of the initial activity is retained by uniformly coupled and randomly coupled N-isopropylacrylamide-enzyme preparations, respectively, after 180 min incubation at 60C
60
-
-
the native enzyme retains 20% of the initial activity after incubation at 60C
63
-
-
thermal unfolding of native SBM occurs at 63C
63.4
-
-
melting temperature of the native enzyme
70
-
P14518
soluble enzyme retains 53% activity after incubation at 70C, immobilized enzyme retains over 70% activity after incubation at 70C
100
-
-
recombinant bromelain shows no activity after 30 min of incubation at 100C
additional information
-
-
thermal denaturation is consistent with an irreversible two-state model
additional information
-
-
immobilization offers more resistance to denaturation particularly at 60C
additional information
-
-
casein stabilizes against heat inactivation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
bromelain immobilized to IDA-Sepharose 6B matrix loaded with Cu2+, Ni2+ and Zn2+ is more resistant to thermal inactivation, as evidenced by retention of over 50% activity after incubation at 60C
-
Casein stabilizes against heat inactivation
-
denaturation in guanidine hydrochloride, the deglycosylated enzyme is more sensitive than the glycosylated one, midpoints of transition are at 2.28 M and 2.86 M, respectively
-
enzymatic activity of bromelain remains uninfluenced by the immobilization of heparin on it
-
enzyme affinity-bound to a Sepharose matrix precoupled with the lactin concanavalin A is more stable to thermal inactivation than native enzyme or enzyme covalently coupled to the CNBr-activated Sepharose
-
glutaraldehyde-crosslinked bromelain has comparable activity to the native enzyme, is more stable against urea, guanidine hydrochloride and temperature-induced inactivation and exhibit better storage ability compared to the unmodified protease
-
immobilization on amino-Sepharose leads to higher proteolytic activity and remarkably enhanced thermal stability as compared to soluble bromelain and that coupled to CNBr- activated Sepharose
P14518
increase in ionic strength by addition of salts results in folded structures somewhat different from the native enzyme. Salt-induced intermediates are characterized by increase in helical content and a significantly reduced exposure of hydrophobic clusters relative to the state at pH 2.0. Salt-induced state shows non-cooperative thermal denaturation alcohol-induced intermediates of the enzyme exhibit increased helical content. Alcohol-induced state shows a cooperative thermal transition
-
Lyophilization causes 27% loss of activity
-
poly(ethylene glycol)-400-induced state has characteristics of molten globule, higher molecular weight poly(ethylene glycol)s cause unfolding of the acid unfolded state
-
Stem bromelain covalently coupled to a thermosensitive polymer of N-isopropylacrylamide either through the amino groups of the enzyme (randomly coupled) or via the lone oligosaccharide chain (uniformly coupled) shows better thermostability. The enzyme coupled via the oligosaccharide chain exhibits better access to the substrate casein as compared to the preparation in which the amino groups formed the point of contact between the enzyme and the polymer.
-
the glycosylated enzyme is more stable than the deglycosylated one
-
the low bromelain activity, particularly in the upper portion of the murine gastrointestinal tract, following oral administration, suggests that extensive gastric inactivation of bromelain may occur in vivo. When formulated in antacid, oral bromelain retains substantial proteolytic activity throughout the gastrointestinal tract (of mice)
-
the pyromellitic anhydride- and poly(maleic anhydride)-modified bromelain does not lose its activity significantly
-
treatment with 10-30% (v/v) 1,1,1,3,3,3-hexafluoroisopropanol induces the partially folded intermediate to adopt much of the native proteins secondary structure, but only a rudimentary tertiary structure, characteristic of the molten globule state. Addition of slightly higher concentrations of 1,1,1,3,3,3-hexafluoroisopropanol causes transformation from an alpha-helix to a beta-sheet and induces formation of a compact nonnative structure
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2,2,2-trifluoroethanol
-
the enhanced binding of 1-anilino-8-naphthalene sulfonic acid to the specific/pre-molten globule (SMG) state of stem bromelain upon addition of 30% 2,2,2-trifluoroethanol suggests the presence of a large number of solvent-accessible non-polar clusters in the treated SMG. The formation of an molten globe (MG)-like state characterized by disordered side chain interactions but with considerable secondary structure when the specific/pre-molten globule (SMG) state of stem bromelain is subjected to 30% 2,2,2-trifluoroethanol (TFE). The TFE-induced MG conformation at alkaline pH could represent the conformation that allows stem bromelain to traverse membranes
Ethanol
-
20% v/v, 37C, 20 min, 33% loss of activity
Ethanol
-
enzyme stabilized in 60-70% ethanol is non-functional, but retains all the elements of secondary structure
guanidine-HCl
-
6 M guanidine-HCl denatured state of stem bromelain is enzymatically inactive
guanidine-HCl
-
activity-enhanced state in the presence of 2 M guanidine-HCl
Methanol
-
25% v/v, 25C, 20 min, stable
urea
-
activity-enhanced state in the presence of 5 M and 8-9 M urea
urea
-
stem bromelain is fully resistant against urea around neutral pH (5.5 to 10.0) and unfolds only below pH 5.0
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
oxidation of the active site thiol leads to the corresponding sulfinic acid, which is catalytically inactive, irreversible oxidative degradation may lead to reduced activity of isozyme SBA/b with substrate Pyr-Phe-Lys-4-nitroanilide
-
653021
photosensitized oxidation of stem bromelain in the presence of methylene blue results in partial loss of the enzymatic activity even if the essential sulfhydryl group is protected against oxidation
-
30274, 30286
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, stored dry, stable
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Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
2 enzyme forms with pI of 9.45 (basic form) and pI 4.7 (acidic form)
-
2 forms: SB1 and SB2
-
2 isozymes SBA/a and SBA/b
-
6 enzyme forms: SBB 1-5 and SBA
-
affinity chromatography
-
bromelain A and B
-
bromelain from pineapple stem and skin are recovered by a PEG 4000/phosphate aqueous two-phase systems (ATPs) liquid-liquid extraction
-
CM-52 cellulose column chromatography
-
enzyme forms: I-1, I-2, II-1, II-2
-
native enzyme from stem, reversible inactivation of the protease in 10 mM Na-phosphate, pH 7.5 containing 5 mM of Na-tetrathionate, during purification, reactivation by 8 mM of DTT and 4 mM of EDTA in the same buffer at 25C, followed by gel filtration
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to homogeneity
-
using Ni-NTA chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cloned into the pENTR/TEV/D-TOPO vector, then sub-cloned into the pDEST17 expression vector. Expression in Escherichia coli BL21
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APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
the complex between stem bromelain and polyclonal Fab* monomer can serve as a model with potential in vivo and industrial application especially in homogenous reaction phases and development of subunit vaccines owing to preponderance of hydrophobic interaction and better diffusion potential of Fab' than of intact IgG
medicine
-
oral bromelain may potentially modify inflammation within the gastrointestinal tract via local proteolytic activity within the colonic microenvironment
medicine
-
bromelain possesses anti-inflammatory activity and reduces blood viscosity, prevents the aggregation of blood platelets, and improves ischemia-reperfusion injury, I/R, in a skeletal muscle model in adult Sprague-Dawley rats. The enzyme increases phosphorylation of Akt in rat heart both in the cytosolic and the nuclear fraction following I/R; stem bromelain triggers an Akt-dependent survival pathway in the heart, revealing a mechanism of cardioprotective action and a potential therapeutic target against ischemia/perfusion injury
medicine
-
bromelain has antitumoral activity
medicine
-
bromelain shows antitumorigenic potential by inducing apoptosis-related proteins along with inhibition of nuclear factor-kappa B driven cyclooxygenase-2 expression by blocking the mitogen-activated protein kinase and Akt/protein kinase B signaling in 7,12-dimethylbenz(a)anthracene-12-O-tetradecanoylphorbol-13-acetate-induced mouse skin tumors
medicine
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bromelain from pineapple stem shows therapeutic benefits in a variety of inflammatory diseases, including murine inflammatory bowel disease, mechanism, overview. Bromelain primary long-term effect is abrogation of firm adhesion of leukocytes to blood vessels at the site of inflammation. These changes in adhesion are correlated with rapid re-expression of the bromelain-sensitive CD62L/L-selectin molecules that mediate rolling following in vivo bromelain treatment and minimal re-expression of CD128
medicine
-
effects of orally administered bromelain in an ovalbumin-induced murine model of acute allergic airway disease in female C57BL/6J mice, bromelain causes decreased methacholine sensitivity, reduction in bronchoalveolar lavage eosinophils and interleukin-13 concentrations, as well as of CD19+ B cells and CD8+ T cells, as compared with controls, overview. Bromelain modulates lung lymphocytes during acute asthma
medicine
-
bromelain has anti-cancer activity, as well as antithrombotic, fibrinolytic, antiedematous, and burn debridement properties
medicine
-
the nonnative form of SBM shows only a fraction of the native protein's enzymatic activity but is more effective at inhibiting cell growth and displays better antitumorigenic properties than native SBM